1. Tweak Your Technique Electrophysiology 101 August 19, 2011 Kevin Lee

2. What can be studied with electrophysiology? Single channel behaviour – in expression systems: HEK293, CHO’s, X. Laevis oocytes Intrinsic properties of excitable cells – neurons, myocytes Synaptic transmission – neuromuscular junction, neuronal synapses

4. The hippocampus

5. Field recordings

6. LTP/LTD

7. Whole-cell Recordings

8. Patch-Recording Methods

9. Whole-cell recordings Current-clamp vs Voltage-clamp “EPSP” “EPSC”

10. Whole-cell recordings Current-clamp vs Voltage-clamp

11. Voltage-clamp

12. mEPSCs (minis) mEPSC amplitude reflects synaptic AMPAR content Beique et al. (PNAS, 2006)

13. mEPSCs (minis) mEPSC frequency can reflect: - total number of synapses - presynaptic function

14. Release probability (Pr) of glutamate Changes in frequency of mEPSCs is generally believed to reflect changes in Pr. “Paired-Pulse” electrical stimulation can also provide information about Pr. Beique et al. (PNAS, 2006)

15. What about Molecular interventions?

16. Biolistic transfection of cortical neurons in organotypic slices Beique & Andrade (J. Physiol, 2002; cover photo)

17. Biolistic transfection of cortical neurons in organotypic slices

18. Barria & Malinow (Neuron, 2002)Cull-Candy et al. (Science STKE, 2004)

19. Drawbacks to electrical stimulation: - Difficult to isolate individual synaptic events - Spatial Parameters left unknown

20. 2P-Uncaging of MNI-Glutamate

21. Béïque et al., PNAS 2006 Uncaging of MNI-Glu activates both AMPA and NMDA receptors High spatial resolution of glutamate uncaging

22. Single Synapse Ca 2+ Imaging Time  1 2 1 2 Uncage Simultaneous Glutamate Uncaging and Calcium Imaging

23. Fundamental dendritic properties probed by glutamate uncaging and calcium imaging Losonczy & Magee (Neuron, 2006)

24. Fundamental dendritic properties probed by glutamate uncaging and calcium imaging Losonczy & Magee (Neuron, 2006)

25. Experimental Wizardry to Envy Kravitz et al. (Nature, 2009)

26. Experimental Wizardry to Envy Kravitz et al. (Nature, 2009)

27. Multiple Simultaneous Recordings in Slices Olah et al. (Nature, 2009)

28. The End